Methods for preparing oltipraz

ABSTRACT

The invention provides improved methods of synthesizing oltipraz, which result in higher overall yield and better purity of the desired product.

RELATED APPLICATIONS

This application claims the benefit of priority to Indian provisional patent application serial number 1891/DEL/2015, filed Jun. 25, 2015; the contents of which are hereby incorporated by reference.

BACKGROUND

Oltipraz, 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, depicted in Formula I, below, was originally developed as an anti-schistosomal drug that eliminates parasitic worms. Later, it was discovered that oltipraz is a chemopreventive agent. The drug also has other known and unknown uses in the medical field.

Various syntheses of oltipraz are known in the art. However, each suffers from various disadvantages, such as low overall yield, long reaction times, risk of explosion due to the use of hydride anion, and severe environmental contamination from required large excesses of P₂S₅.

There exists a need for a fast, safe method of synthesizing oltipraz in high overall yield without using large excesses of P₂S₅.

SUMMARY

In certain embodiments, the invention relates to a method comprising the steps of:

-   -   a) combining in a first container a first solvent and a first         base;     -   b) stirring the contents of the first container at a temperature         of about 0° C. for about 5 minutes;     -   c) adding to the first container, over a period of time of about         15 minutes, methyl propionate;     -   d) stirring the contents of the first container at about 0° C.         for about 15 minutes;     -   e) adding to the first container a solution of

in a second solvent, wherein the solution is added over a period of time of about 30 minutes while warming the first container to a temperature of about 22° C.; and the second solvent is a mixture of tetrahydrofuran and 1,4-dioxane; and

-   -   f) stirring the contents of the first container at about 22° C.         for a first period of time.

In certain embodiments, the invention relates to any of the methods described herein, wherein the first base is sodium pentanoate or potassium t-butoxide.

In certain embodiments, the invention relates to any of the methods described herein, wherein the method is a method of synthesizing

in a yield greater than 80%, greater than 81%, greater than 82%, greater than 83%, greater than 84%, greater than 85%, greater than 86%, greater than 87%, greater than 88%, greater than 89%, greater than 90%, greater than 91%, greater than 92%, greater than 93%, greater than 94%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or greater than 99%.

In certain embodiments, the invention relates to a method of synthesizing oltipraz, comprising the steps of:

-   -   i) combining in a second container P₂S₅ and a first quantity of         toluene;     -   ii) heating the second container at a temperature of about 100°         C.;     -   iii) adding to the second container a solution of

in a second quantity of toluene; and

-   -   iv) heating the contents of the second container to reflux for a         second period of time,

wherein no xylene is added or included at any step.

In certain embodiments, the invention relates to any of the methods described herein, wherein the purity of the oltipraz produced by the claimed methods is greater than 97%, greater than 98%, or greater than 99%, as determined by gas chromatography.

In certain embodiments, the invention relates to any of the methods described herein, wherein the overall yield of oltipraz from

is greater than 21%, greater than 22%, greater than 23%, greater than 24%, greater than 25%, greater than 26%, greater than 27%, greater than 28%, greater than 29%, or greater than 30%.

In certain embodiments, the invention relates to any of the methods described herein, wherein the overall yield of oltipraz from

is greater than 21%, and the purity of the resulting oltipraz is greater than 97%, as determined by gas chromatography.

DETAILED DESCRIPTION I. Overview

In certain embodiments, the invention relates to an improved method of synthesizing oltipraz. In certain embodiments, the method involves Step 2 or Step 3 (or both Step 2 and Step 3 in succession) as depicted in Scheme 1.

In certain embodiments, the invention relates to any one of the methods described herein, wherein Step 2 does not involve sodium hydride.

In certain embodiments, the invention relates to any one of the methods described herein, wherein the yield of Step 2 is greater than 80%, greater than 81%, greater than 82%, greater than 83%, greater than 84%, greater than 85%, greater than 86%, greater than 87%, greater than 88%, greater than 89%, greater than 90%, greater than 91%, greater than 92%, greater than 93%, greater than 94%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or greater than 99%.

In certain embodiments, the invention relates to any one of the methods described herein, wherein the overall yield of Step 2 and Step 3 is greater than 21%, greater than 22%, greater than 23%, greater than 24%, greater than 25%, greater than 26%, greater than 27%, greater than 28%, greater than 29%, or greater than 30%.

In certain embodiments, the invention relates to any one of the methods described herein, wherein the purity of the oltipraz produced by the claimed methods is greater than 97%, greater than 98%, or greater than 99%, as determined by gas chromatography.

In certain embodiments, the invention relates to any one of the methods described herein, wherein the overall yield of Step 2 and Step 3 is greater than 21%, and the purity of the resulting oltipraz is greater than 97%, as determined by gas chromatography.

II. Exemplary Methods

It will be understood that any reaction described herein, in any of its variations, can be combined in sequence with one or more of the other reactions described herein, in any of their variations, substantially in analogy with the sequence shown in Scheme 1.

In certain embodiments, the invention relates to a method comprising the steps of:

-   -   a) combining in a first container a first solvent and a first         base;     -   b) stirring the contents of the first container at a temperature         of about 0° C. for about 5 minutes;     -   c) adding to the first container, over a period of time of about         15 minutes, methyl propionate;     -   d) stirring the contents of the first container at about 0° C.         for about 15 minutes;     -   e) adding to the first container a solution of

in a second solvent, wherein the solution is added over a period of time of about 30 minutes while warming the first container to a temperature of about 22° C.; and the second solvent is a mixture of tetrahydrofuran and 1,4-dioxane; and

-   -   f) stirring the contents of the first container at about 22° C.         for a first period of time.

In certain embodiments, the invention relates to any of the methods described herein, wherein the first solvent is tetrahydrofuran.

In certain embodiments, the invention relates to any of the methods described herein, wherein the first solvent is a mixture of tetrahydrofuran and 1,4-dioxane. In certain embodiments, the invention relates to any of the methods described herein, wherein the first solvent is about a 5:1, about a 4:1, or about a 3:1 mixture by volume of tetrahydrofuran and 1,4-dioxane. In certain embodiments, the invention relates to any of the methods described herein, wherein the first solvent is about a 4:1 mixture by volume of tetrahydrofuran and 1,4-dioxane.

In certain embodiments, the invention relates to any of the methods described herein, wherein the first base is sodium pentanoate or potassium t-butoxide.

In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of the first base in the first solvent is from about 1.0 M to about 1.8 M.

In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of the first base in the first solvent is about 1.0 M, about 1.2 M, about 1.4 M, about 1.6 M, or about 1.8 M. In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of the first base in the first solvent is about 1.4 M.

In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of first base to methyl propionate is from about 3:1 to about 1:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of first base to methyl propionate is about 3:1, about 2:1, or about 1:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of first base to methyl propionate is about 1:1.

In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of

in the second solvent is from about 1.6 M to about 2.0 M. In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of

in the second solvent is about 1.6 M, about 1.8 M, or about 2.0 M. In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of

in the second solvent is about 1.8 M.

In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of methyl propionate to

is from about 2:1 to about 1.6:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of methyl propionate to

is about 2:1, about 1.8:1, or about 1.6:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of methyl propionate to

is about 1.8:1.

In certain embodiments, the invention relates to any of the methods described herein, wherein the second solvent is from about a 1.2:1 to about a 0.8:1 mixture by volume of tetrahydrofuran and 1,4-dioxane. In certain embodiments, the invention relates to any of the methods described herein, wherein the second solvent is about a 1.2:1, about a 1:1, or about a 0.8:1 mixture by volume of tetrahydrofuran and 1,4-dioxane. In certain embodiments, the invention relates to any of the methods described herein, wherein the second solvent is about a 1:1 mixture by volume of tetrahydrofuran and 1,4-dioxane.

In certain embodiments, the invention relates to any of the methods described herein, wherein the first period of time is from about 2 h to about 10 h. In certain embodiments, the invention relates to any of the methods described herein, wherein the first period of time is about 2 h, about 3 h, about 4 h, about 5 h, about 6 h, about 7 h, about 8 h, about 9 h, or about 10 h. In certain embodiments, the invention relates to any of the methods described herein, wherein the first period of time is about 6 h.

In certain embodiments, the invention relates to any of the methods described herein, wherein the method is a method of synthesizing

In certain embodiments, the invention relates to any of the methods described herein, wherein the method is a method of synthesizing

in a yield greater than 80%, greater than 81%, greater than 82%, greater than 83%, greater than 84%, greater than 85%, greater than 86%, greater than 87%, greater than 88%, greater than 89%, greater than 90%, greater than 91%, greater than 92%, greater than 93%, greater than 94%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or greater than 99%.

In certain embodiments, the invention relates to any of the methods described herein, wherein the method is a method of synthesizing

in a yield greater than 80%, greater than 81%, greater than 82%, greater than 83%, greater than 84%, greater than 85%, greater than 86%, greater than 87%, greater than 88%, greater than 89%, greater than 90%, greater than 91%, greater than 92%, greater than 93%, greater than 94%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or greater than 99% without purification other than isolation.

In certain embodiments, the invention relates to any of the methods described herein, wherein the method consists essentially of steps (a)-(f).

In certain embodiments, the invention relates to a method comprising the steps of:

-   -   i) combining in a second container P₂S₅ and a first quantity of         toluene;     -   ii) heating the second container at a temperature of about 100°         C.;     -   iii) adding to the second container a solution of

in a second quantity of toluene; and

-   -   iv) heating the contents of the second container to reflux for a         second period of time, wherein no xylene is added or included at         any step.

In certain embodiments, the invention relates to a method comprising the steps of:

-   -   i) combining in a second container P₂S₅, a first quantity of         toluene, a first quantity of water, and a phase transfer         catalyst;     -   ii) heating the second container at a temperature of about 100°         C.;     -   iii) adding to the second container a solution of

in a second quantity of toluene; and

-   -   iv) heating the contents of the second container to reflux for a         second period of time,

wherein no xylene is added or included at any step.

In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of P₂S₅ in the first quantity of toluene is from about 0.2 M to about 0.5 M. In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of P₂S₅ in the first quantity of toluene is about 0.2 M, about 0.25 M, about 0.3 M, about 0.35 M, about 0.4 M, about 0.45 M, or about 0.5 M.

In certain embodiments, the invention relates to any of the methods described herein, wherein P₂S₅ and the first quantity of toluene are combined in the second container under an inert atmosphere. In certain embodiments, the invention relates to any of the methods described herein, wherein P₂S₅ and the first quantity of toluene are combined in the second container under a nitrogen atmosphere.

In certain embodiments, the invention relates to any of the methods described herein, wherein the volume ratio of the first quantity of toluene to the first quantity of water is about 12:1, about 10:1, or about 8:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the volume ratio of the first quantity of toluene to the first quantity of water is about 10:1.

In certain embodiments, the invention relates to any of the methods described herein, wherein the phase transfer catalyst is a tetraalkylammonium salt or a tetraalkylphosphonium salt. In certain embodiments, the invention relates to any of the methods described herein, wherein the phase transfer catalyst is a tetrabutylphosphonium salt. In certain embodiments, the invention relates to any of the methods described herein, wherein the phase transfer catalyst is tetrabutylphosphonium halide. In certain embodiments, the invention relates to any of the methods described herein, wherein the phase transfer catalyst is tetrabutylphosphonium chloride.

In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of

in the second quantity of toluene is from about 0.2 M to about 0.4 M. In certain embodiments, the invention relates to any of the methods described herein, wherein the concentration of

in the second quantity of toluene is about 0.2 M, about 0.3 M, or about 0.4 M.

In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of P₂S₅ to

is from about 1.2:1 to about 0.6:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of P₂S₅ to

is about 1.2:1, about 1.1:1, about 1:1, about 0.9:1, about 0.8:1, about 0.7:1, or about 0.6:1.

In certain embodiments, the invention relates to any of the methods described herein, wherein the second period of time is from about 2 h to about 10 h. In certain embodiments, the invention relates to any of the methods described herein, wherein the second period of time is about 2 h, about 3 h, about 4 h, about 5 h, about 6 h, about 7 h, about 8 h, about 9 h, or about 10 h. In certain embodiments, the invention relates to any of the methods described herein, wherein the second period of time is about 6 h.

In certain embodiments, the invention relates to any of the methods described herein, wherein the

is synthesized according to steps (a)-(f), described above.

In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of

to P₂S₅ is from about 0.8:1 to about 1.8:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of

to P₂S₅ is about 0.8:1, about 0.9:1, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 1.6:1, about 1.7:1, or about 1.8:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of

to P₂S₅ is about 0.9:1. In certain embodiments, the invention relates to any of the methods described herein, wherein the mole ratio of

to P₂S₅ is about 1.4:1.

In certain embodiments, the invention relates to any of the methods described herein, wherein the method is a method of synthesizing oltipraz.

In certain embodiments, the invention relates to any of the methods described herein, wherein the overall yield of oltipraz from

is greater than 21%, greater than 22%, greater than 23%, greater than 24%, greater than 25%, greater than 26%, greater than 27%, greater than 28%, greater than 29%, or greater than 30%.

In certain embodiments, the invention relates to any of the methods described herein, wherein the purity of the oltipraz produced by the claimed methods is greater than 97%, greater than 98%, or greater than 99%, as determined by gas chromatography.

In certain embodiments, the invention relates to any of the methods described herein, wherein the overall yield of oltipraz from

is greater than 21%, and the purity of the resulting oltipraz is greater than 97%, as determined by gas chromatography.

In certain embodiments, the invention relates to any of the methods described herein, wherein the method consists essentially of steps (i)-(iv).

In certain embodiments, the invention relates to any of the methods described herein, wherein the method consists essentially of steps (a)-(f) and steps (i)-(iv).

In certain embodiments, the invention relates to any one of the methods described herein, further comprising the steps outlined in any other method described herein.

In certain embodiments, the invention relates to the use of any one of the compounds described herein in the manufacture of a medicament.

Definitions of variables in the structures in the schemes herein are commensurate with those of corresponding positions in the formulae delineated herein.

In certain embodiments, the compounds described herein contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids. Optical isomers may be prepared, for example, by resolving a racemic mixture. The resolution can be carried out in the presence of a resolving agent, by chromatography or by repeated crystallization or by some combination of these techniques which are known to those skilled in the art. Further details regarding resolutions can be found in Jacques, et al., Enantiomers. Racemates, and Resolutions (John Wiley & Sons, 1981).

The synthesized compounds can be separated from a reaction mixture and further purified by a method such as column chromatography, high pressure liquid chromatography, or recrystallization. As can be appreciated by the skilled artisan, further methods of synthesizing the compounds of the formulae herein will be evident to those of ordinary skill in the art. Additionally, the various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. In addition, the solvents, temperatures, reaction durations, etc. delineated herein are for purposes of illustration only and one of ordinary skill in the art will recognize that variation of the reaction conditions can produce the desired bridged macrocyclic products of the present invention. Synthetic chemistry transformations useful in synthesizing the compounds described herein are known in the art and include, for example, those such as described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons (1991): L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and subsequent editions thereof.

The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

EXEMPLIFICATION

The present invention is further illustrated by the following Example which should not be construed as limiting in any way. The Examples and discoveries described herein are representative. As such, the studies and results described in the Examples section herein may be used as a guideline.

Example 1—Synthesis of Pyrazine-2-Carboxylic Acid Methyl Ester, ST-601 (Step 1)

Input:

Pyrazinoic acid (OT-1) (1758 g, SM)

Sulfuric Acid (69.5 g) MeOH (8.8 L) Procedure:

-   -   Charged MeOH (8.8 L) and OT-1 (1758 g) at RT.     -   Charged H₂SO₄ (69.5 g) in one portion (21-22° C. exotherm).         Heated to (60-65° C.) and stirred at 55-65° C. for 19 hours.

19 h, 96.0% OT-2 and 4.0% OT-1 by HPLC

-   -   Reaction was cooled to 15/30° C. No precipitate formed.     -   NaHCO₃ (180 g) was charged in lots. The solution bubbled         slightly and quickly went from yellow to pink. The mixture was         stirred for 5 min at 15/30° C.     -   The mixture was then concentrated to 1.5-2.5 vol at ≤30° C.     -   Charged NaCl (0.700 g) in water (2.5 vol). Upon stirring,         solution became clear.     -   Stirred for 15 min at 15/30° C. After stirring, the aqueous         layer became slightly cloudy. The solids were filtered off and         the layers were separated.     -   The aqueous layer was extracted with DCM (3×2 vol.). TLC         indicated that extraction was complete after 3rd extraction.     -   Organic layers were dried over anhydrous Na₂SO₄ (0.4 g/g SM).     -   Concentrated to 1.5-2.5 vol under vacuum at ≤30° C. Some         precipitate formed.     -   Charged heptanes (8 vol) over a minimum of 30 min. Pale white         slurry. Let stir overnight.     -   Stirred at −5/−15° C. for a minimum of 1 hr. Solids were         filtered off and rinsed with cold heptanes     -   (2×1 vol.) Pulled solids dry on filter for 10 min. Dried in         vacuum oven at ≤30° C. to constant mass. Dried over weekend.         Solids went from pale white to light brown.     -   Color changed, but no degradation was observed.         Output material: ST-601

Lot No.: 2463-24-1

Appearance: light Brown Solids

Yield: 1721 g (91.0%)

HPLC purity: 98.9% ¹H NMR—Conforms to structure

Example 2—Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Input: ST-601 (1000 g); Lot #2463-24-1

Methyl propionate (64 g+893 g); Lot #11-2713-56+11-2713-57+R11-1913-101 NaH in mineral oil, 60 wt. %, (579 g) Toluene (10 vol)

Procedure:

-   -   Charged ST-601 (1 kg), NaH, 60 wt. % (579 g) and toluene (10         vol) and stirred at 15/25° C. No reaction (no gas evolution,         exotherm or appearance change).     -   Charged methyl propionate (64 g). No immediate reaction (no gas         evolution, exotherm or appearance change) at 15/25° C.     -   Charged MeOH (85 g) at 15/25° C. Immediately started to react         (gas evolution, exotherm). Heated to 30/40° C.     -   Charged methyl propionate (893 g) at 30/40° C. over 5 h. The         reaction was slower in the beginning, but became faster (more         exotherm and gas evolution) after 1-2 h when ˜0.3 eq of methyl         propionate was charged. Stirred at 30/40° C. for 88 h.         64 h, 30/35° C., brown slurry, practically no gas evolution         observed, IPC HPLC1: 74.4%+14.1%=88.5% of ST-602 at 4.81 min and         5.62 min; 1.2% of ST-601 at 1.97 min; 5.8% of “Int” at 3.21 min.

72 h, 35° C., IPC HPLC2: 47.8%+42.9%=90.7% of ST-602; 0.9% of ST-601; 4.3% of “Int” 88 h, 38° C., IPC HPLC3: 60.3%+32.6%=92.9% of ST-602; 0.64% of ST-601; 1.8% of “Int”

-   -   Reaction was cooled to 15/20° C.     -   Charged AcOH (2.5 eq) over a minimum of 1 hr. Exothermic.         Slightly thick, brown suspension.     -   Charged 10% NaCl solution (8 vol) over a minimum of 1 h at         15/30° C. Slight exotherm. Brown, biphasic solution. Let stir at         15/30° C. for a minimum of 30 min to dissolve all solids.     -   The phases were separated. Both the aqueous and the organic         phases were brown.     -   The organic phase was washed with a 10% NaCl solution (5 vol).     -   The organic phase was washed with NaHCO₃ (0.1 g/g SM) in 10% aq.         NaCl solution (5 vol.)     -   The organic phase was dried over anhydrous sodium sulfate (0.2         g/g SM) for a minimum of 2 hrs.     -   Solids were filtered off and rinsed with Toluene (1×1 vol)     -   Concentrated to 3-4 vol at ≤50° C.         Isolated material: ST-602

Lot No.: 2463-52-2

Appearance: light brown liquid Yield: Assumed 100% (1406 g net directly used in next step)

HPLC: 94.3% of ST-602 Example 3—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Input:

ST-602 (from 1 kg of ST-601 (SM), 1 eq); Lot #2463-52-2 P₂S₅ (1931 g); Lot #11-1513-52 Toluene (15 vol)

Observations:

-   -   Charged ST-602 in toluene (˜3.5 L; 3-4 vol) to P₂S₅ (1931 g) in         toluene (10 L; 10 vol). Rinsed with toluene (1 L; 1 vol). No         exothermic effect, yellow slurry.     -   Reaction over after 36 h at 95-97° C.

36 h, −95° C., IPC HPLC lot #2463-54-1: 95.2% of ST-617 at 6.85 min and 4.8% of ST-602 at 4.79 min

-   -   Work-up (after 36 h at −95° C.): Cooled to RT over 20 mins     -   Charged the slurry via a transfer line to a mixture of Na₂CO₃         (2686 g; 3.5 eq), water (15 L; 15 vol) and THF (5 L; 5 vol) at         15/30° C. over a minimum of 1 h—weak exothermic effect and no         gas evolution observed.     -   The resulting mixture was slowly transferred back to the parent         flask—some gas evolution observed. Minor insolubles were         observed.     -   The resulting mixture was stirred at RT for 65 h (shouldn't need         more than 12 h) to complete gas evolution/quench of the reaction         mixture     -   The resulting mixture was passed through a 0.5″ celite bed to         remove fine insolubles.     -   The organic layer was separated, dried with Na₂SO₄ (200 g; 0.2         g/g SM) and concentrated under reduced pressure at ≤50° C. to         2-4 L (2-4 vol) residue volume (3 L actual)—product precipitates         during concentration.     -   The resulting slurry was diluted with MeOH (1.5 L; 1.5 vol) and         stirred at RT for 2 h.     -   The solids were collected by filtration, rinsed with MeOH (2×500         mL; 2×0.5 vol) followed by heptane (1 L+0.5 L; 1+0.5 vol).     -   Dried in air to yield constant weight.         Isolated material: ST-603 (crude ST-617)

Lot No.: 2463-55-1

Appearance: red solid powder (crystals not uniform)

Yield: 223 g (14%)

HPLC: 99% by R&D method

Example 4—Purification of Crude ST-617 (Step 4) Input: ST-603 (220 g); Lot #2463-55-1 DMSO (2.2 L+220 mL)

Water (330 mL; 1.5 vol)

Observations:

-   -   Added ST-603 into DMSO (2.2 L) and heated to 65° C.     -   Clear solution in DMSO at ˜65° C. Heated further to ˜80° C. and         hot filtered at ˜80° C.     -   Hot filtration was easy to perform (no fast precipitation of         product).     -   Diluted with water at 70/85° C. to initiate product         precipitation.     -   The mixture was chilled/stirred at RT for 1 hour     -   Collected the solids by filtration, rinsed with 5:1 DMSO/water         (2×1.5 vol) and MeOH (2×1.5 vol) sequentially.     -   Re-slurried in MeOH (8 mL) at RT for 70 h (shouldn't need more         than 4-6 hours) to help wash out DMSO and collected by         filtration.     -   Finally rinsed with MeOH (2×1.5 vol) and dried in a vacuum oven         at 30/40° C. to yield constant weight.         Isolated material: ST-617

Lot No.: 2463-57-1

Appearance: brown-red solid

Yield: 191 g (87%)

HPLC: 99.5% by R&D method ¹H NMR (CDCl₃): conforms to structure (clean) with residual DMSO (0.3 wt. %)

Example 5—Synthesis of Pyrazine-2-Carboxylic Acid Methyl Ester, ST-601 (Step 1)

Aim: Carry out 170-g-scale Step 1

Procedure:

-   1. Added 900 mL of MeOH and 175 g of OT-1 into a 2-neck 2-liter RBF     at 22° C. on a heating mantle. -   2. Added 7 grams of concentrated sulfuric acid in one portion into     the RBF. -   3. Heated to 65° C. and stirred for 24 hours. -   4. TLC at 24 hrs indicates little to no OT-1 (Rf 0.1 in 95:5     Hexane:EtOAc) left and new spot which is the same as authentic     ST-601 at Rf 0.7 -   5. Reaction was cooled to RT (22° C.). No precipitate formed. -   6. 75 g of Sodium Bicarbonate was added. Slight gas evolution and     colour goes from yellow to pink. -   7. The mixture was stirred for 10 min at 22° C. -   8. The mixture was transferred to a 1-neck RBF and set on a rotovap. -   9. The mixture was then concentrated to ˜200 mL at ≤30° C. -   10 Added a prepared solution of 70 g of NaCl in 400 mL of water.     Upon stirring, the solution became clear. -   11 Stirred for 10 min at 22° C. After stirring, the solution became     slightly cloudy. The solids were filtered on a Buchner filter. -   12. The aqueous layer was extracted three times with 200 ml of     dichloromethane each time -   13. The dichloromethane layers were combined and 50 g of anhydrous     sodium sulfate was added to the solution. Stirred for 5 min and     filtered on Buchner. -   14. The solution was concentrated to ˜50 mL on a Rotovap under     vacuum at ≤30° C. Some precipitate was visible but very fine. -   15. Added 700 mL of mixed heptanes over ˜45 min. A pale white slurry     formed which was stirred gently overnight. -   16. Cooled the RBF to ˜15° C. (dry ice batch) and stirred for 2     hours. -   17. Thicker solids formed which were filtered on a Buchner. -   18. The residue was rinsed with refrigerated mixed heptanes two     times with 200 mL each. -   19. Pulled the solids dry on the Buchner for 15 min. Transferred to     tared dish. -   20. Set to dry in vacuum oven at ≤30° C. overnight. -   21. Color of powder is pale brown. -   22. GC (in lab) shows 98.4% purity versus authentic -   23. Weight of powder is 168 g (89% yield) -   24. Labelled as ST-601-001

Example 6—Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Establish baseline for Step 2 using NaH as the base.

Procedure

-   1. Add 2.5 g (0.0181 mole) of ST-601, 1.4 g of 60% NaH in oil and 25     mL of toluene into a 100-mL 2-neck RBF at 22° C. and stir for 30 min     to thoroughly strip the oil off the NaH. No gas evolution or     temperature change was observed. -   2. Added 250 mg of methyl propionate into the flask with no exotherm     or gas evolution observed. -   3. Added 2.5 mL of methanol. -   4. Immediately started to react with gas evolution and exotherm. -   5. Heated to 40° C. -   6. Added 2.3 g of methyl propionate dropwise from addition funnel at     1 mL/20 min. -   7 The reaction became more vigorous after 30 min of addition. -   8. Stirred at 40° C. -   9. Monitored by HPLC (GeoChem Method ST1). After 24 hrs at 40° C.     the reaction is a brown slurry and no gas was being evolved. HPLC     showed two peaks (same as for authentic ST-602) totaling 82.4% and     2.1% of ST-601 and with an intermediate peak of 15.2% (all area %     peaks) -   10. After 72 h at 40° C. the reaction is a brown slurry with no gas     being evolved. HPLC showed two peaks (same as for authentic ST-602)     totaling 96.0% and 0.5% of ST-601 with an intermediate peak of 1%     (all area % peaks) -   11. The reaction was cooled to 22° C. (RT) and 3 mL of glacial     acetic acid was added slowly. (some slow gas evolution). Then added     in 100 mL of 10% aq. NaCl solution with stirring. The organic layer     was separated and washed with 100 mL of 10% aq. NaCl+10% NaHCO₃ and     dried over 10 g of sodium sulfate anhydrous. The slurry was filtered     on a Buchner and the reside washed 1× with 20 mL of toluene. -   12. Concentrated on a rotovap at ≤50° C. to ˜5 mL. -   13. The above reference standard reaction product will be considered     the crude standard required for use of un-isolated ST-602 in the     next cyclization step. -   14. The orange-brown solution will be used as is in Step 3     cyclization yield verification step. -   15. Labeled as ST-602-002

Example 7—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with t-butoxide

Procedure

-   1. Added 4.1 g (0.0362 mole) of potassium t-butoxide powder and 25     mL of dry THF into a 100-mL 2-neck RBF at 0° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turns yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF and     added into the reaction over 30 minutes while allowing the reaction     to come to RT (˜½ hr). No gas evolution or exotherm. -   5. Stirred at reflux for 24 hours -   6. Monitored by HPLC (Method ST1). After 24 hrs at reflux the     reaction is a brown slurry -   7. HPLC showed two peaks (same as for authentic ST-602) totaling     32.6% and 39.7% of ST-601 and with an intermediate peak of 18% (all     area % peaks) -   8. After 72 hrs at reflux the reaction was a dark brown slurry. -   9. HPLC showed two peaks (same as for authentic ST-602) totaling     36.8% and 38.2% of ST-601 with an intermediate peak of 12.5% (all     area % peaks) -   10. Concentrated on a rotovap at ˜50° C. -   11. The 2.5 dark brown solution will be used as is in Step 3     cyclization yield verification step. -   12. Labeled as ST-602-003

Example 8—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with sodium pentanoate

Procedure

-   1. Add 4.5 g (0.0362 mole) of sodium pentanoate (Mwt 124) powder and     25 mL of dry THF into a 100-mL 2-neck RBF at 0° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. Add 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF     and added into the reaction over 30 minutes while allowing the     reaction to come to RT (˜½ hr). No gas evolution or exotherm. -   5. Stirred at reflux for 24 hours -   6. Monitored by HPLC (Method ST1). After 24 hrs at reflux the     reaction is a yellow-brown slurry -   7. HPLC showed two peaks (same as for authentic ST-602) totaling     18.3% and 20.4% of ST-601 and with an intermediate peak of 39.9%     (all area % peaks) -   8. After 72 hrs at reflux the reaction is a dark brown slurry. -   10. HPLC showed two peaks (same as for authentic ST-602) totaling     26.9% and 21.7% of ST-601 with an intermediate peak of 35.3% (all     area % peaks) -   11. Concentrated on a rotovap at ≤50° C. to ˜5 mL -   12. The dark brown solution will be used as is in Step 3 cyclization     yield verification step. -   13. Labeled as ST-602-004

Example 9—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Establish baseline for Step 3

Procedure

-   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL of Toluene into a     100-mL 2-neck RBF under nitrogen -   2. Yellow slurry but easily stirable. -   3. Added crude ST-602-002 solution (˜5 mL). Yellow-brown slurry. -   4. Started stirring and heated to 50° C., color changed to reddish     brown in 10 min. Slow H₂S gas evolution was observed which is passed     through a caustic bubbler and no H₂S was exited. -   5. Continued heating up to 95° C. and let stir for 36 hours. -   6. A sample was taken for HPLC (method ST-1). No starting material     present. Product ST-603 was present but also many other small peaks.     Area % of ST-603 is 88.8% against authentic sample with 5.4% ST-602 -   7. The reaction was cooled to 22° C. (RT) -   8. In a 100-mL beaker was prepared a quenching mixture of 7 g of     solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT. -   9. To the above beaker was slowly added (over 10 min) the content of     the reaction flask. Weak exotherm was observed but no H₂S gas was     smelt at all. -   10. The resulting mixture was stirred over 12 hours to complete     quench of the reaction (in a very high vent hood attached to a     scrubber for safety). Some insolubles were visible at the end of the     12 hours. -   11. The reaction slurry was passed through a 1″-celite bed to remove     fine insolubles. -   12. The organic layer was separated (both are colored red), dried     over 2 g anhydrous sodium sulfate, and concentrated on a rotovap at     ≤50° C. to 20 mL volume. Solid ppt is observed during concentration. -   13. The resulting slurry was diluted with 20 mL of methanol and     stirred at RT for 2 h. -   14. The solids were collected by filtration, rinsed 2× with 20 mL of     refrigerated cold methanol followed by 20 mL of cold heptane. -   15. Dried in vacuum oven to constant weight. -   16. Weight of powder is 348 mg (˜12% yield) -   17. HPLC showed 98.2% product against authentic sample. -   18. Sent 20 mg to IPAC for ¹H NMR. -   19. Labelled as ST-603-005

Example 10—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 to study cyclization of crude mixture from potassium t-butoxide condensation.

Procedure

-   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL of Toluene into a     100-mL 2-neck RBF under nitrogen. -   2. Yellow slurry. -   3. Added crude ST-602-003 solution (˜5 mL). Brown slurry (difficult     to stir) -   4. Started stirring and heated to 50° C. No color change. Slow H₂S     gas evolution is observed which is passed through a caustic bubbler     and no H₂S is exited. -   5. Continued heating up to 95° C. and let stir for 36 hours. -   6. A sample was taken for HPLC (method ST-1): 601 (25%), and 602     (18%), and 8% ST-603 was present, but also many other small peaks. -   7. The reaction was cooled to 22° C. (RT) -   8. In a 100-mL beaker was prepared a quenching mixture of 7 g of     solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT. -   9. To the above beaker was slowly added (over 10 min) the content of     the reaction flask. Weak exotherm was observed but strong smell of     H₂S gas. -   10. The resulting mixture was stirred over 12 hours to completely     quench the reaction (in a very high vent hood attached to a scrubber     for safety). Lots of insolubles are visible at the end of the 12     hours. -   11. The reaction slurry was passed through a 1″ celite bed to remove     fine insolubles. Very difficult filtration, so left for over 12     hours. -   12. After finally getting two phase filtrate, the organic layer was     separated (both are colored brown), dried over 2 g anhydrous sodium     sulfate and concentrated on a rotovap at ≤50° C. to 20 mL volume. No     solid ppt was observed during concentration. -   13. The resulting solution was diluted with 20 mL of methanol and     stirred at RT for 2 h. -   14. No solids were observed, so added 20 mL of cold heptane. Still     no solids. Left in refrigerator for 24 hrs but still no solids     formed.

Example 11—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 to study cyclization of crude mixture from sodium pentanoate condensation.

Procedure

-   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL of Toluene into a     100-mL 2-neck RBF under nitrogen. -   2. Yellow slurry. -   3. Added crude ST-602-003 solution (˜5 mL). Brown stirrable slurry -   4. Started stirring and heated to 50° C. Slight color change to     reddish hue. Slow H₂S gas evolution is observed which is passed     through a caustic bubbler and no H₂S is exited. -   5. Continued heating up to 95° C. and let stir for 36 hours. -   6. A sample was taken for HPLC (method ST-1): 601 (10%) and 602     (17%) and 43% ST-603 was present, but also many other small peaks. -   7. The reaction was cooled to 22° C. (RT) -   8. In a 100-mL beaker was prepared a quenching mixture of 7 g of     solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT. -   9. To the above beaker was slowly added (over 10 min) the content of     the reaction flask. Weak exotherm was observed and some smell of H₂S     gas. -   10. The resulting mixture was stirred over 12 hours to completely     quench the reaction (in a very high vent hood attached to a scrubber     for safety). Some insolubles are visible at the end of the 12 hours. -   11. The reaction slurry was passed through a 1″ celite bed to remove     fine insolubles. Difficult filtration over 6 hour period. -   12. Two phase filtrate both red/brown in color—the organic layer was     separated, dried over 2 g anhydrous sodium sulfate and concentrated     on a rotovap at ≤50° C. to 20 mL volume. No solid ppt was observed     during concentration. -   13. The resulting solution was diluted with 20 mL of methanol and     stirred at RT for 2 h. -   14. No solids were observed, so added 20 mL of cold heptane. Still     no solids. Left in refrigerator for 24 hrs, but still no solids     formed.

Example 12—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with excess sodium pentanoate

Procedure

-   1. Add 9 g (0.0724 mole) of sodium pentanoate (MWt 124) powder and     25 mL of dry THF into a 100-mL 2-neck RBF at 0° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. Add 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF     and added into the reaction over 30 minutes while allowing the     reaction to come to RT (˜½, hr). No gas evolution or exotherm. -   5. Stirred at reflux for 24 hours -   6. Monitored by HPLC (Method ST1). After 24 hrs at reflux the     reaction is a yellow-brown slurry -   7. HPLC showed two peaks for ST-602 totaling 72.4%, and 12.1% of     ST-601 and with an intermediate peak of 8.3% (all area % peaks) and     many small peaks that were not present in baseline experiment -   8. After 72 hrs at reflux the reaction is a dark brown slurry. -   9. HPLC shows two peaks for ST-602 totaling 77.5%, and 9.2% of     ST-601 with an intermediate peak of 4.1% (all area % peaks) and same     smaller peaks. -   10. Concentrated on a rotovap at ≤50° C. to ˜5 mL -   11. The dark brown solution may be used as is in P₂S₅ cyclization     step. -   12. Labeled as ST-602-008

Example 13—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 to study cyclization of crude mixture from sodium pentanoate condensation.

Procedure

-   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL of Toluene into a     100-mL 2-neck RBF under nitrogen. -   2. Thick, yellow slurry. -   3. Added crude ST-602-003 solution (˜5 mL). Brown, stirrable slurry -   4. Started stirring and heated to 50° C. Slight color change to     red-brown. Slow H₂S gas evolution was observed, which was passed     through a caustic bubbler and no H₂S was exited. -   5. Continued heating up to 95° C. and let stir for 36 hours. -   6. A sample was taken for HPLC (method ST-1): 601 (10%) and 602     (17%) and 43% ST-603 was present, but also many other small peaks. -   7. The reaction was cooled to 22° C. (RT) -   8. In a 100-mL beaker was prepared a quenching mixture of 7 g of     solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT. -   9. To the above beaker was slowly added (over 10 min) the content of     the reaction flask. Weak exotherm was observed and some smell of H₂S     gas. -   10. The resulting mixture was stirred over 12 hours to completely     quench the reaction (in a very high vent hood attached to a scrubber     for safety). Some insolubles were visible at the end of the 12     hours. -   11. The reaction slurry was passed through a 1″ celite bed to remove     fine insolubles. Difficult filtration over 6 hour period. -   12. Two phase filtrate both red/brown in color—the organic layer was     separated, dried over 2 g anhydrous sodium sulfate, and concentrated     on a rotovap at ≤50° C. to 20 mL volume. No solid ppt was observed     during concentration. -   13. The resulting solution was diluted with 20 mL of methanol and     stirred at RT for 2 h. -   14. No solids were observed, so added 20 mL of cold heptane. Still     no solids. Left in refrigerator for 24 hrs and some solids were     visible. -   15. Reduced the volume to 5 mL on a rotovap and added fresh cold 20     mL heptane. Left in refrigerator overnight. -   16. The solids were collected by filtration, rinsed 2× with 20 mL of     refrigerated cold methanol followed by 20 mL of cold heptane. -   18. Dried in vacuum oven to constant weight. -   19. Weight of Powder is 139 mg (˜4.8% yield) -   20. HPLC shows 94.8% ST-617 product against authentic sample. -   21. Labelled as ST-603-009

Example 14—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate

Procedure

-   1. Add 4.5 g (0.0362 mole) of sodium pentanoate (MWt 124) powder and     25 mL of dry THF into a 100-mL 2-neck RBF at 0° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF and     added into the reaction over 30 minutes while allowing the reaction     to come to RT (˜½ hr). No gas evolution or exotherm. -   5. Stirred for 24 hours at RT -   6. TLC shows product (ST-602) spot (Rf 0.4) begins forming after 2     hrs at RT and an intermediate spot (Rf 0.2) and spot for ST-601 is     still also visible. Second 2 hr TLC shows more product spot but     still shows both intermediate and ST-601 spots. After 6 hrs ST-602     spot and only trace of intermediate and ST-601. Stirred overnight.     After 16 hrs no change from 6 hr TLC. -   7. 20 mL of distilled water and 20 mL of saturated sodium chloride     solution were added to the reaction solution and stirred for 30     minutes. -   8. The reaction solution was concentrated to a volume of 40 mL and     then extracted with 2×25 mL of toluene. -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered -   10. The filtrate was rotovaped to give 1.9 g crude ST-602 as a dark     brown viscous oil -   11. Labeled as ST-602-010

Example 15—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate and vary temperature

Procedure

-   1. Add 4.5 g (0.0362 mole) of sodium pentanoate (Mwt 124) powder and     25 mL of dry THF into a 100-mL 2-neck RBF at 0° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. Add 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF     and added into the reaction over 30 minutes while allowing the     reaction to come to RT (˜½ hr). No gas evolution or exotherm. -   5. Warmed to 40° C. -   6. Monitored on TLC every 2 hours -   7. TLC showed product (ST-602) spot (Rf 0.4) after 2 hrs at RT with     almost no ST-601 but a long streaking along the plate. -   8. Continued at 40° C. to complete the experiment for 6 hrs -   9. 20 mL of distilled water and 20 mL of saturated sodium chloride     solution were added to the reaction solution and stirred for 30     minutes. -   10. The reaction solution was concentrated to a volume of 45 mL and     then extracted with 2×25 mL of toluene -   11. The resultant extract was almost black in color and had     suspended solids and was very tarry in form -   15. Labeled as ST-602-011 but kept for discard.

Example 16—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate, use warmer temperature for formation of anion, and stop reaction after 6 h

Procedure

-   1. Add 4.5 g (0.0362 mole) of sodium pentanoate (MWt 124) powder and     25 mL of dry THF into a 100-mL 2-neck RBF at 10° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (MWt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 10° C. -   4. Add 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF     and added into the reaction over 30 minutes while allowing the     reaction to come to RT (˜½ hr). No gas evolution or exotherm. -   5. Stirred at RT for 6 hours. Monitor on TLC every 2 hours -   6. TLC showed product (ST-602) spot (Rf 0.4) begins forming after 2     hrs at RT and an intermediate spot (Rf 0.2) and spot for ST-601 is     still also visible. Second 2 hr TLC showed more product spot but     still showed both intermediate and ST-601 spots. After 6 hrs ST-602     spot and only trace of intermediate and ST-601 is visible. -   7. 20 mL of distilled water and 20 mL of saturated sodium chloride     solution were added to the reaction solution and stirred for 30     minutes -   8. The reaction solution was concentrated to a volume of 45 mL and     then extracted with 2×25 mL of toluene. -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered. -   10. The filtrate was rotovaped to give 2.7 g crude ST-602 as a brown     oil with no tarry nature. -   11. Labeled as ST-602-012 and carried forward for cyclization     experiment

Example 17—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 using the solvent system described in U.S. Pat. No. 7,288,652 to establish baseline

Procedure

-   1. Added 5.65 g (0.025 mole) of P₂S₅ and 25 mL Toluene+25 mL mixed     xylenes into a 100-mL 2-neck RBF under nitrogen. -   2. Yellow slurry but easily stirrable. -   3. Heated to 120° C. while stirring -   4. Added crude ST-602-012 solution (˜10 mL in toluene). Yellow-brown     slurry. -   5. Started stirring and heated to reflux (˜140° C.). Color changes     to reddish brown in 5 mins. Slow H₂S gas evolution was observed,     which was passed through a caustic bubbler and no H₂S is exited. -   6. Continued reflux for 6 hours. -   7 A sample was taken for HPLC (method ST-1): no starting material     present, and product ST-603 was present but also some other small     peaks. -   8. The reaction was cooled to 25° C. (RT) -   9. In a 100-mL beaker was prepared a quenching mixture of 7 g of     solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT. -   10. To the above beaker was slowly added (over 10 min) the content     of the reaction flask. Weak exotherm was observed but no H₂S gas     smell at all. -   11. The resulting mixture was stirred over 3 hours to complete     quench of the reaction (in a very high vent hood attached to a     scrubber for safety). Very few insolubles are visible at the end of     the 3 hours. -   12. The reaction slurry was passed through a 1″ celite bed to remove     the fine insolubles. -   13. The organic layer was separated (both are colored red), dried     over 2 g anhydrous sodium sulfate, and concentrated on a rotovap at     ≤50° C. to 20 mL volume. -   14. The resulting slurry was diluted with 20 mL of methanol and     stirred at RT for 2 h resulting in solids -   15. The solids were collected by filtration, rinsed 2× with 20 mL of     refrigerated cold methanol followed by 20 mL of cold heptane. -   16. Dried in vacuum oven to constant weight. -   17. Weight of crude ST-603 crystalline powder is 609 mg (˜21% yield) -   18. Labelled as ST-603-013.

Example 18—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate, use 15% 1,4-dioxane in THF as solvent system for reaction

Procedure

-   1. Added 4.5 g (0.0362 mole) of sodium pentanoate (MWt 124) powder     and 25 mL of dry THF into a 100-mL 2-neck RBF at 0° C. Stirred for 5     minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turns yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 5 mL of THF+5 mL     of 1,4-Dioxane and added into the reaction over 30 minutes while     allowing the reaction to come to RT (˜½ hr). No gas evolution or     exotherm. -   5. Stirred at RT for 6 hours. Monitored by TLC every 2 hours. -   6. TLC shows product (ST-602) spot (Rf 0.4) begins forming after 2     hrs at RT and an intermediate spot (Rf 0.2) and spot for ST-601 is     still also visible. Second 2 hr TLC shows more product spot but     still shows both intermediate and ST-601 spots. After 6 hrs, ST-602     spot and only trace of intermediate and ST-601 spots were visible. -   7. 30 mL of distilled water and 30 mL of saturated sodium chloride     solution were added to the reaction solution (extra aqueous layer     added due to more polar 1,4-dioxane and stirred for 30 minutes. -   8. The reaction solution was concentrated to a volume of 60 mL and     then extracted with 2×25 mL of toluene. -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered. -   10. The filtrate was rotovaped to give 3.1 g crude ST-602 as a brown     oil with no tarry nature -   11. Labeled as ST-602-014 (GC shows 84% ST-002 peak)

Example 19—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate, use 1,4-dioxane as solvent system for reaction

Procedure

-   1. Added 4.5 g (0.0362 mole) of sodium pentanoate (Mwt 124) powder     and 25 mL of dry 1,4-dioxane into a 100-mL 2-neck RBF at 0° C.     Stirred for 5 minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of     1,4-Dioxane and added into the reaction over 30 minutes while     allowing the reaction to come to RT (˜½ hr). No gas evolution or     exotherm. -   5. Stirred at RT for 6 hours. Monitored by TLC every 2 hours. -   6. TLC showed product (ST-602) spot (Rf 0.4) begins forming after 2     hrs at RT but multiple new trailing spots also present. After 6 hrs,     ST-602 spot, no ST-601 but lots of multiple smaller spots were     visible. -   7. 50 mL of distilled water and 50 mL of saturated sodium chloride     solution were added to the reaction solution (extra aqueous layer     added due to more polar 1,4-dioxane) and stirred for 30 minutes. -   8. The reaction solution was concentrated to a volume of 100 mL     (using a high vacuum on the rotovap) and then extracted with 2×25 mL     of toluene -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered -   10. The filtrate was rotovaped to give 1.8 g crude ST-602 as a brown     oil with some tarry nature -   11. Labeled as ST-602-015 (GC shows 68% ST-002 peak)

Example 20—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate, use 1,4-dioxane as solvent system for reaction, complete reaction at 0° C.

Procedure

-   1. Added 4.5 g (0.0362 mole) of sodium pentanoate (MWt 124) powder     and 25 mL of dry 1,4-dioxane into a 100-mL 2-neck RBF at 0° C.     Stirred for 5 minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned yellow. -   3. Stirred for an additional 15 minutes at 0° C. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of     1,4-Dioxane and added into the reaction over 30 minutes. No gas     evolution or exotherm. -   5. Stirred at 0° C. for 6 hours. Monitored by TLC every 2 hours. -   6. TLC shows a small amount of product (ST-602) spot (Rf 0.4) began     forming after 2 hrs at RT. Intermediate spot was visible and large     ST-601 spot. After 4 hrs, no real change in TLC. Skipped the 6 hr     TLC and stirred overnight (12 hrs) at 0° C. TLC now shows more     ST-602 but still ˜20% ST-601 was visible. Allowed the reaction to     warm up to 10° C. and stirred for 2 hrs more. All the ST-601 was     gone, many trialing spots were visible so reaction was taken for     workup. -   7. 50 mL of distilled water and 50 mL of saturated sodium chloride     solution were added to the reaction solution (extra aqueous layer     added due to more polar 1,4-dioxane) and stirred for 30 minutes -   8. The reaction solution was concentrated to a volume of 110 mL     (using a high vacuum on the rotovap) and then extracted with 2×25 mL     of toluene. -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered -   10. The filtrate was rotovaped to give 2.0 g crude ST-602 as a brown     oil with almost no tarry nature. -   11. Labeled as ST-602-016 (GC showed 71% ST-002 peak)

Example 21—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× sodium pentanoate, use 30% 1,4-dioxane in THF as solvent system for reaction

Procedure

-   1. Added 4.5 g (0.0362 mole) of sodium pentanoate (MWt 124) powder     and 20 mL of dry THF+5 mL 1,4-dioxane into a 100-mL 2-neck RBF at     0° C. Stirred for 5 minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned pale     yellow. -   3. Stirred for an additional 15 minutes at 0° C. No color change. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 5 mL of THF+5 mL     of 1,4-Dioxane and added into the reaction over 30 minutes while     allowing the reaction to come to RT (˜½ hr). No gas evolution or     exotherm. -   5. Stirred at RT for 6 hours. Monitored by TLC every 2 hours. -   6. TLC showed ˜60% product (ST-602) spot (Rf 0.4) began forming     after 2 hrs at RT and an intermediate spot (Rf 0.2) and <10% spot     for ST-601 was still also visible. Second 2 hr TLC showed more     product spot and intermediate spot but no ST-601 spot. After 6 hrs     ST-602 spot only and some trailing spots visible. -   30 mL of distilled water and 30 mL of saturated sodium chloride     solution were added to the reaction solution and stirred for 30     minutes -   8. The reaction solution was concentrated to a volume of 65 mL     (vacuum of 14 mmHg was adequate on rotovap) and then extracted with     2×30 mL of toluene -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered. -   10. The filtrate was rotovaped to give 3.3 g crude ST-602 (MWt=194)     as a brown oil with no tarry nature -   11. Labeled as ST-602-017 to be carried forward for cyclization     experiment

Example 22—Alternative Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× potassium t-butoxide, use 30% 1,4-dioxane in THF as solvent system for reaction

Procedure

-   1. Added 4.1 g (0.0362 mole) of potassium t-butoxide powder     (MWt 112) powder and 20 mL of dry THF+5 mL 1,4-dioxane into a 100-mL     2-neck RBF at 0° C. Stirred for 5 minutes -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into the     flask using a dropping funnel over 15 minutes. Color turned pale     yellow. -   3. Stirred for an additional 15 minutes at 0° C. No color change. -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 5 mL of THF+5 mL     of 1,4-Dioxane and added into the reaction over 30 minutes while     allowing the reaction to come to RT (˜½ hr). No gas evolution or     exotherm. -   5. Stirred at RT for 6 hours. Monitor on TLC every 2 hours. -   6. Very similar TLC profile as previous Example. TLC showed ˜60%     product (ST-602) spot (Rf 0.4) began forming after 2 hrs at RT and     an Intermediate spot (Rf 0.2) and <10% spot for ST-601 was still     also visible. Second 2 hr TLC showed more product spot and     intermediate spot but no ST-601 spot. -   30 mL of distilled water and 30 mL of saturated sodium chloride     solution were added to the reaction solution and stirred for 30     minutes. -   8. The reaction solution was concentrated to a volume of 65 mL     (vacuum of 14 mmHg was adequate on rotovap) and then extracted with     2×30 mL of toluene. -   9. The resultant extract was dried over anhydrous magnesium sulfate     and filtered. -   10. The filtrate was rotovaped to give 3.1 g crude ST-602 (MWt=194)     as a brown oil with no tarry nature. -   11. Labeled as ST-602-018 will be carried forward for cyclization     experiment

Example 23—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 to study cyclization of much cleaner crude mixture from condensation.

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL Toluene into a         100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Heated to 100° C. while stirring     -   4. Added crude ST-602-019a solution (12.4 g solution in         toluene). Yellow-brown slurry.     -   5. Started stirring and heated to reflux (˜110° C.). Color         changed to reddish brown in 15 mins. Slow H₂S gas evolution was         observed which is passed through a caustic bubbler and no H₂S is         exited.     -   6. Continued reflux for 6 hours.     -   7. A sample was taken for HPLC (method ST-1). No starting         material present. Product ST-603 was present but also some other         small peaks.     -   8. The reaction was cooled to 25° C. (RT)     -   9. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   10. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. Weak exotherm was observed but         no H₂S gas smell at all.     -   11. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Very few insolubles are visible at the end         of the 3 hours.     -   12. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Much more easily filtered than         comparative ST-603-005.     -   13. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   14. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in solids     -   15. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol followed by 20 mL of cold         heptane.     -   16. Dried in vacuum oven to constant weight.     -   17. Weight of crude ST-603 crystalline powder is 874 mg (˜21%         yield) (Mol Wt=226) GC=97.3%     -   18. Labelled as ST-603-020.

Example 24—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 to study cyclization of much cleaner crude mixture from condensation using solvent system and slightly higher reflux temperature from U.S. Pat. No. 7,288,652

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 25 mL toluene+30 mL         xylene into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirred.     -   3. Heated to 130° C. while stirring     -   4. Added crude ST-602-019b solution (˜10 mL in toluene).         Yellow-brown slurry.     -   5. Started stirring and heated to reflux (˜110° C.). Color         changed to reddish brown in 15 mins. Slow H₂S gas evolution is         observed which is passed through a caustic bubbler and no H₂S is         exited.     -   6. Continued reflux for 6 hours.     -   7. A sample was taken for HPLC (method ST-1). No starting         material present. Product ST-603 is present but also some other         small peaks.     -   8. The reaction was cooled to 25° C. (RT)     -   9. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   10. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. Weak exotherm was observed but         no H₂S gas smell at all.     -   11. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Very few insolubles are visible at the end         of the 3 hours.     -   12. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. easy filtration may even work with         paper.     -   13. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   14. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in solids     -   15. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol followed by 20 mL of cold         heptane.     -   16. Dried in vacuum oven to constant weight.     -   17. Weight of crude ST-603 crystalline powder is 826 mg—very         similar and within error of the yield produced by the method of         the previous Example, so apparently the higher boiling point         does not materially affect to the reaction yield.     -   18. Labelled as ST-603-021.

Example 25—Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2)

Aim: Replace base in Step 2 with 2× potassium t-butoxide, as described in U.S. Pat. No. 7,288,652

Procedure

-   -   1. Added 4.1 g (0.0362 mole) of potassium t-butoxide powder         (MWt 112) powder and 25 mL of dry THF into a 100-mL 2-neck RBF         at 0° C. Stirred for 5 minutes     -   2. Added 2.9 g (0.0326 mole) of methyl propionate (Mwt 88) into         the flask using a dropping funnel over 15 minutes. Color turned         pale yellow.     -   3. Stirred for an additional 15 minutes at 0° C. No color         change.     -   4. 2.5 g (0.0181 mole) of ST-601 was dissolved in 10 mL of THF         and added into the reaction over 30 minutes while allowing the         reaction to come to RT (˜½ hr). No gas evolution or exotherm.     -   5. Stirred at RT for 6 hours. Monitored by TLC every 2 hours.     -   6. TLC shows ˜60% product (ST-602) spot (Rf 0.4) began forming         after 2 hrs at RT and an intermediate spot (Rf 0.2) and <10%         spot for ST-601 was still also visible. Second 2 hr TLC showed         more product spot and intermediate spot but no ST-601 spot.     -   7. 30 mL of distilled water and 30 mL of saturated sodium         chloride solution were added to the reaction solution and         stirred for 30 minutes.     -   8. The reaction solution was concentrated to a volume of 65 mL         (vacuum of 14 mmHg was adequate on rotovap) and then extracted         with 2×30 mL of toluene.     -   9. The resultant extract was dried over anhydrous magnesium         sulfate and filtered.     -   10. The filtrate was rotovaped to give 2.7 g crude ST-602         (Mwt=194) as a brown oil little tarry nature.     -   11. Labeled as ST-602-022 will be carried forward for         cyclization experiment as per U.S. Pat. No. 7,288,652 for total         yield comparisons.

Example 26—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 to study overall yield an purity of Steps 1, 2, and 3 as described in U.S. Pat. No. 7,288,652

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 25 mL toluene+30 mL         xylene into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirred.     -   3. Heated to 130° C. while stirring 4. Added crude ST-602-019b         solution (˜10 mL in toluene). Yellow-brown slurry.     -   5. Started stirring and heated to reflux (˜110° C.). Color         changed to reddish brown in 15 mins. Slow H₂S gas evolution was         observed which is passed through a caustic bubbler and no H₂S is         exited.     -   6. Continued reflux for 6 hours.     -   7. A sample was taken for HPLC (method ST-1). No starting         material present. Product ST-603 was present but also some other         small peaks.     -   8. The reaction was cooled to 25° C. (RT)     -   9. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   10. To the above beaker was slowly added (over 10 min) the         content of the reaction flask. Weak exotherm was observed but no         H₂S gas smell at all.     -   11. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Very few insolubles are visible at the end         of the 3 hours.     -   12. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. easy filtration may even work with         paper.     -   13. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   14. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in solids     -   15. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol followed by 20 mL of cold         heptane.     -   16. Dried in vacuum oven to constant weight.     -   17. Weight of crude ST-603 crystalline powder was 708 mg.         Labelled as ST-603-023. GC shows purity of 96.7%.

Example 27—Catalyzed Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 using tetrabutyl phosphonium chloride in toluene+water

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL Toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of tetrabutyl phosphonium chloride     -   4. Heated to 100° C. while stirring     -   5. Added crude ST-602-034a solution (12 g solution in toluene).         Yellow-Green slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to red-brown after 20 min. Some H₂S gas evolution was         observed which was passed through a caustic bubbler and no H₂S         was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. ˜50% starting material present.         Product ST-603 is also present but some other spots also         present.     -   9. Continued reflux for 2 hours.     -   10. A sample was taken for TLC. <5% starting material present.         Product ST-603 is also present but some other spots also         present.     -   11. The reaction was cooled to 25° C. (RT)     -   12. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   13. To the above beaker was slowly added (over 10 min) the         content of the reaction flask. No exotherm was observed and no         H₂S gas smell at all.     -   14. The resulting mixture was stirred for 2 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Some insolubles are visible at the end of         the 2 hours.     -   15. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   16. The organic layer was separated (both were colored         red-brown), dried over 2 g anhydrous sodium sulfate and         concentrated on a rotovap at ≤50° C. to 20 mL volume.     -   17. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in sticky solids only but         titutration with hexane did yield approx. 400 mg of flowable         red-brown powder. TLC shows mostly ST-603—sample saved as         ST-603-35.

Example 28—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene under reflux pressure

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene into a         200 mL SS316 autoclave under nitrogen.     -   2. Added crude ST-602-019d solution (˜10 mL in toluene).         Yellow-brown slurry.     -   3. Started stirring, sealed the autoclave, and heated to jacket         temperature 180° C.     -   4. Pressure gauge showed increasing pressure from 1 ATM to 16.4         ATM after 2 hours and held steady.     -   5. Continued 2 additional hours for a total of 4 hours.     -   6. Released the pressure into a caustic bubbler.     -   7. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   8. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. Weak exotherm was observed and         some H₂S gas could be detected by smell.     -   9. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Few insolubles were visible at the end of         the 3 hours.     -   10. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles.     -   11. The organic layer was separated (both layers were colored         red), dried over 2 g anhydrous sodium sulfate, and concentrated         on a rotovap at ≤50° C. to 20 mL volume.     -   12. The resulting slurry was diluted with 20 mL of methanol and         stirred at about 23° C. (room temperature or RT) for 2 h,         resulting in solids.     -   13. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol, followed by 20 mL of cold         heptane.     -   14. Dried in vacuum oven to constant weight.     -   15. Weight of crude ST-603 brown pasty powder is 943 mg. TLC         showed the product spot but many other smaller spots. Labelled         as ST-603-024. GC purity=64.3%.

Example 29—Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2) Aim: Scale-up Step 2 Procedure

-   -   1. Add 32.8 g (0.2896 mole) of potassium t-butoxide powder         powder and 160 mL of dry THF+40 mL 1,4-dioxane into a 500-mL         2-neck RBF at 0° C. Stirred for 5 minutes     -   2. Added 23.2 g (0.2608 mole) of methyl propionate into the         flask using a dropping funnel over 15 minutes. Color turned pale         yellow.     -   3. Stirred for an additional 15 minutes at 0° C. No color         change.     -   4. Added 20 g (0.1448 mole) of ST-601 dissolved in 40 mL of         THF+40 mL of 1,4-dioxane over 30 minutes while allowing the         reaction to come to RT. No gas evolution or exotherm was         observed.     -   5. Stirred at RT for 12 hours. Monitored by TLC every 2 hours.     -   6. 240 mL of distilled water and 240 mL of saturated sodium         chloride solution were added to the reaction solution, which was         subsequently stirred for 30 minutes.     -   7. The reaction solution was concentrated to a volume of 510 mL         (vacuum of H mmHg was adequate on rotovap) and then extracted         with 2×240 mL of toluene.     -   8. The resultant extract was rotovaped to give 97.7 g crude         ST-602 as a brown oil with no tarry nature.     -   9. The solution was divided into eight equal parts and labelled         as ST-602-25a, b, d, e f, g, h for comparative cyclization         experiments.

Example 30—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene in autoclave under reflux

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene into a         200-mL SS316 autoclave under nitrogen.     -   2. Added crude ST-602-025a solution (˜10 mL in toluene).         Yellow-brown slurry.     -   3. Started stirring, sealed the autoclave, and heated to jacket         temperature 180° C.     -   4. Pressure gauge showed increasing pressure from 1 ATM to 16.1         ATM after 2 hours and held steady.     -   5. Released the pressure into a caustic bubbler.     -   6. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   7. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. A weak exotherm was observed and         some H₂S gas was detected.     -   8. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Few insolubles were visible at the end of         the 3 hours.     -   9. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles.     -   10. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   11. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h, resulting in solids     -   12. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol, followed by 20 mL of cold         heptane.     -   13. Dried in vacuum oven to constant weight.     -   14. Weight of crude ST-603 brown pasty powder was 802 mg. TLC         showed some product but also many less concentrated side         products. Labelled as ST-603-026.

Example 31—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene in autoclave under reflux for 12 h

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene into a         200-mL SS316 autoclave under nitrogen.     -   2. Added crude ST-602-025b solution HO mL in toluene).         Yellow-brown slurry.     -   3. Started stirring, sealed the autoclave, and heated to jacket         temperature 180° C.     -   4. Pressure gauge showed increasing pressure from 1 ATM to 16.1         ATM after 2 hours and held steady.     -   5. Held under pressure overnight.     -   6. Released the pressure into a caustic bubbler.     -   7. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   8. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. A weak exotherm was observed and         some H₂S gas was detected.     -   9. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Few insolubles were visible at the end of         the 3 hours.     -   10. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles.     -   11. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   12. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in solids.     -   13. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol followed by 20 mL of cold         heptane.     -   14. Dried in vacuum oven to constant weight.     -   15. Weight of crude ST-603 brown pasty powder was 985 mg. TLC         showed some product but many less concentrated side products         were also visible. Labelled as ST-603-027. GC purity=54%.

Example 32—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in xylene in autoclave under reflux

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL xylene into a         200-mL SS316 autoclave under nitrogen.     -   2. Added crude ST-602-025c solution HO mL in toluene).         Yellow-brown slurry.     -   3. Started stirring, sealed the autoclave and heated to jacket         temperature 180° C.     -   4. Pressure gauge showed increasing pressure from 1 ATM to 15.1         ATM after 2 hours and held steady.     -   5. Continued 2 additional hours for a total of 4 hours. 6.         Released the pressure into a caustic bubbler.     -   7. In a 100-mL beaker was prepared a quenching mixture of 7 g of         solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   8. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. A weak exotherm was observed and         some H₂S gas was detected.     -   9. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Few insolubles were visible at the end of         the 3 hours.     -   10. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles.     -   11. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   12. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in solids.     -   13. The solids were collected by filtration, rinsed 2× with 20         mL of refrigerated cold methanol followed by 20 mL of cold         heptane.     -   14. Dried in vacuum oven to constant weight.     -   15. Weight of crude ST-603 brown pasty powder was 764 mg. TLC         showed some product but also many less concentrated side         products. Labelled as ST-603-028. GC purity=68%.

Example 33—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using an ammonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of Aliquat 336         (N-methyl-N,N,N-trioctan-1-ammonium chloride).     -   4. Heated to 100° C. while stirring     -   5. Added crude ST-602-025d solution (12.4 g solution in         toluene). Yellow slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to reddish brown in 15 mins. Fast H₂S gas evolution was         observed; gas was passed through a caustic bubbler and no H₂S         was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. No starting material present.         Product ST-603 was present but also many other small peaks.     -   9. The reaction was cooled to 25° C.     -   10. In a 100-ml beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   11. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. A weak exotherm was observed but         no H₂S gas was detected.     -   12. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Very few insolubles were visible at the         end of the 3 hours.     -   13. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   14. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   15. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in sticky solids only—no clean         powder resulted, so reaction was discarded.

Example 34—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using an ammonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of benzalkonium chloride.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-025e solution (12.4 g solution in         toluene). Yellow slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to dark brown in 5 mins. Very fast H₂S gas evolution was         observed; the gas was passed through a caustic bubbler and no         H₂S was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. No starting material present.         Very little product ST-603 was present and lots of streaking was         visible on the TLC.     -   9. With the TLC showing poor conversion the reaction was         discarded.

Example 35—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using an ammonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of benzyltrimethylammonium tribromide.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-025f solution (12.4 g solution in         toluene). Brown slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changes to brown in 15 min. Moderate H₂S gas evolution was         observed; the gas was passed through a caustic bubbler and no         H₂S was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. No starting material present.         Product ST-603 was present but also many other compounds in         lower concentrations.     -   9. The reaction was cooled to 25° C.     -   10. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   11. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. A weak exotherm was observed but         no H2s gas was detected.     -   12. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Very few insolubles were visible at the         end of the 3 hours.     -   13. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   14. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   15. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in sticky solids only.     -   16. Titurated the sticky solids with a mixture of DMSO+water         until brown/red solid powder was formed on the side of the         beaker.     -   17. Collected the solids and checked by TLC. ST-603 was present         but still many other trailing spots.     -   18. Reaction was discarded.

Example 36—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using an ammonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of tributylammonium acetate.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-025 g solution (12.4 g solution in         toluene). Greenish slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to greenish red-brown in 10 min. Some H₂S gas evolution         was observed; the gas was passed through a caustic bubbler and         no H₂S was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. Starting material present.         Product ST-603 was also present but a large streak was visible         at bottom of TLC.     -   9. Refluxed for 2 more hours.     -   10. A sample was taken for TLC. No more starting material         present. Product ST-603 was present with a small streak at         bottom of TLC     -   11. The reaction was cooled to 25° C.     -   12. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   13. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. A weak exotherm was observed but         no H₂S gas was detected.     -   14. The resulting mixture was stirred over 3 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Very few insolubles were visible at the         end of the 3 hours.     -   15. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. VERY slow filtration.     -   16. The organic layer was separated (both are colored red),         dried over 2 g anhydrous sodium sulfate, and concentrated on a         rotovap at ≤50° C. to 20 mL volume.     -   17. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in sticky solids only—no clean         powder resulted, so reaction was discarded.

Example 37—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using an ammonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of tetraethylammonium iodide.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-025h solution (12.4 g solution in         toluene). Brown slurry.     -   6. Started stirring and heated to reflux (˜110° C.). No color         change after 20 mins. No H₂S gas evolution was observed.         Regardless, a caustic bubbler was used and no H₂S was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. Mostly starting material present.     -   9. Refluxed for 2 more hours.     -   10. A sample was taken for TLC. Still mostly starting material         present. Faint ST-603 spot was present with streaking at bottom         of TLC.     -   11. Refluxed overnight.     -   12. TLC showed little starting material remaining, but still         only a faint ST-603 spot.     -   13. Reaction was discarded.

Example 38—Synthesis of Methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, ST-602 (Step 2) Aim: Scale-up Step 2 Procedure

-   -   1. Add 32.8 g (0.2896 mole) of potassium t-butoxide powder and         160 mL of dry THF+40 mL 1,4-dioxane into a 500-mL 2-neck RBF at         0° C. Stirred for 5 minutes.     -   2. Added 23.2 g (0.2608 mole) of methyl propionate into the         flask using a dropping funnel over 15 minutes. Color turned pale         yellow.     -   3. Stirred for an additional 15 minutes at 0° C. No color         change.     -   4. Add 20 g (0.1448 mole) of ST-601 was dissolved in 40 mL of         THF+40 mL of 1,4-dioxane and added into the reaction over 30         minutes while allowing the reaction to come to RT (˜½ hr). No         gas evolution or exotherm.     -   5. Stirred at RT for 12 hours. Monitored by TLC every 2 hours.     -   6. 240 mL of distilled water and 240 mL of saturated sodium         chloride solution were added to the reaction solution and         stirred for 30 minutes.     -   7. The reaction solution was concentrated to a volume of 510 mL         (vacuum of 14 mmHg was adequate on rotovap) and then extracted         with 2×240 mL of toluene.     -   8. The resultant extract was rotovaped to give 97.7 g crude         ST-602 as a brown oil with no tarry nature.     -   9. Divided the solution into eight equal parts and labelled as         ST-602-34a, b, c, d, e, f, g, h for comparative cyclization         experiments.

Example 39—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using a phosphonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of tetrabutyl phosphonium chloride.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-034a solution (12 g solution in toluene).         Yellow-green slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to red-brown after 20 min. Some H₂S gas evolution was         observed; gas was passed through a caustic bubbler and no H₂S         was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. ˜50% starting material present.         Product ST-603 was also present, in addition to some side         products.     -   9. Continued reflux for 2 hours.     -   10. A sample was taken for TLC. <5% starting material present.         Product ST-603 was present, in addition to some side products.     -   11. The reaction was cooled to 25° C.     -   12. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   13. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. No exotherm was observed and no         H₂S gas was detected.     -   14. The resulting mixture was stirred for 2 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Some insolubles were visible at the end of         the 2 hours.     -   15. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   16. The organic layer was separated (both are colored         red-brown), dried over 2 g anhydrous sodium sulfate, and         concentrated on a rotovap at ≤50° C. to 20 mL volume.     -   17. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in sticky solids, but         titutration with hexane yielded approximately 400 mg of flowable         red-brown powder. TLC showed mostly ST-603—sample saved as         ST-603-35.

Example 40—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using a phosphonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of methyltriphenoxyphosphonium iodide.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-034b solution (12 g solution in toluene).         Yellow slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to brown in 1 min. Some H₂S gas evolution was observed;         gas was passed through a caustic bubbler and no H₂S was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. No starting material present.         Product ST-603 was present but a large number of other compound         also present.     -   9. The reaction was cooled to 25° C.     -   10. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   11. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. No exotherm was observed and no         H₂S gas was detected.     -   12. The resulting mixture was stirred for 2 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Some insolubles were visible at the end of         the 2 hours.     -   13. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   14. The organic layer was separated (both are colored         red-brown), dried over 2 g anhydrous sodium sulfate, and         concentrated on a rotovap at ≤50° C. to 20 mL volume.     -   15. The resulting slurry was diluted with 20 ml of methanol and         stirred at RT for 2 h resulting in sticky solids—no clean powder         resulted so reaction was discarded.

Example 41—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using a phosphonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-ml 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of tetraphenyl phosphonium chloride.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-034c solution (12 g solution in toluene).         Yellow-green slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to red-brown after 5 min. Some H₂S gas evolution was         observed; gas was passed through a caustic bubbler and no H₂S         was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. ˜10% starting material present.         Product ST-603 was present, in addition to a number of side         products.     -   9. The reaction was cooled to 25° C.     -   10. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   11. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. No exotherm was observed and no         H₂S gas was detected.     -   12. The resulting mixture was stirred for 2 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Some insolubles were visible at the end of         the 2 hours.     -   13. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   14. The organic layer was separated (both are colored         red-brown), dried over 2 g anhydrous sodium sulfate, and         concentrated on a rotovap at ≤50° C. to 20 mL volume.     -   15. The resulting slurry was diluted with 20 ml of methanol and         stirred at RT for 2 h resulting in sticky solids. Tituration did         not yield any flowable powder product—so, reaction was         discarded.

Example 42—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene and water under reflux using a phosphonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene+5 mL         water into a 100-mL 2-neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 100 mg of trihexyltetradecylphosphonium chloride.     -   4. Heated to 100° C. while stirring     -   5. Added crude ST-602-034d solution (12 g solution in toluene).         Brown slurry.     -   6. Started stirring and heated to reflux (˜110° C.). No color         change after 20 mins. Some H₂S gas evolution was observed; gas         was passed through a caustic bubbler and no H₂S was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. No starting material present.         Very little product spot if at all and many side products         detected—so, reaction was discarded without work up.

Example 43—Synthesis of Crude 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione, ST-617 (Step 3)

Aim: Carry out Step 3 in toluene under reflux using an ammonium salt as a phase transfer catalyst

Procedure

-   -   1. Added 5.65 g (0.025 mole) of P₂S₅ and 50 mL toluene into a         100-ml 2 neck RBF under nitrogen.     -   2. Yellow slurry but easily stirrable.     -   3. Added 200 mg of Aliquat HTA-1.     -   4. Heated to 100° C. while stirring.     -   5. Added crude ST-602-034e solution (12 g solution in toluene).         Yellow slurry.     -   6. Started stirring and heated to reflux (˜110° C.). Color         changed to red-brown after 10 min. Some H₂S gas evolution was         observed; gas was passed through a caustic bubbler and no H₂S         was exited.     -   7. Continued reflux for 2 hours.     -   8. A sample was taken for TLC. <10% starting material present.         Product ST-603 was present but few other spots also present.     -   9. Continued reflux for 2 hours.     -   10. A sample was taken for TLC. <5% starting material present.         Product ST-603 was present but some other spots also present.     -   11. The reaction was cooled to 25° C.     -   12. In a 100-mL beaker was prepared a quenching mixture of 7 g         of solid Na₂CO₃ dissolved in 30 mL water and 10 mL of THF at RT.     -   13. To the above beaker was slowly added (over 10 min) the         contents of the reaction flask. Mild exotherm was observed but         no H₂S gas was detected.     -   14. The resulting mixture was stirred for 2 hours to complete         quench of the reaction (in a very high vent hood attached to a         scrubber for safety). Some insolubles were visible at the end of         the 2 hours.     -   15. The reaction slurry was passed through a 1″ celite bed to         remove the fine insolubles. Slow filtration.     -   16. The organic layer was separated (both are colored         red-brown), dried over 2 g anhydrous sodium sulfate, and         concentrated on a rotovap at ≤50° C. to 20 mL volume.     -   17. The resulting slurry was diluted with 20 mL of methanol and         stirred at RT for 2 h resulting in slighty sticky solids.         Titutration with hexane yielded 830 mg of flowable red-brown         powder. TLC showed mostly ST-603—sample saved as ST-603-39.

INCORPORATION BY REFERENCE

The contents of all references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application are hereby expressly incorporated herein in their entireties by reference. Unless otherwise defined, all technical and scientific terms used herein are accorded the meaning commonly known to one with ordinary skill in the art.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims. The contents of all references, patents, and published patent applications, and patent applications cited throughout this application are incorporated herein by reference. 

1-20. (canceled)
 21. A method of making a composition comprising oltipraz, comprising the steps of: (i) reacting a quantity of pyrazine-2-carboxylic acid methyl ester with methyl propionate to form methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, wherein the reaction is carried out in the presence of a base comprising potassium t-butoxide or sodium pentanoate, and a solvent comprising 1,4-dioxane in tetrahydrofuran (THF), wherein the ratio of 1,4-dioxane in the THF is at least about 1:5; (ii) adding at least one aqueous liquid to quench the reaction in Step (i); (iii) adding a nonpolar organic solvent and an ionic salt, thereby forming a composition comprising an aqueous component and an organic component, wherein the organic component comprises the nonpolar solvent and methyl 2-methyl-3-oxo-3-(pyrazin-2-yl) propanoate; (iv) separating the organic component from the aqueous component; (v) reacting the methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate ester in the organic component with P₂S₅ in the presence of a nonpolar organic solvent to form 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione (oltipraz).
 22. A method according to claim 21, wherein the ratio of 1,4-dioxane to THF in Step (i) is from about 1:5 to about 1:3.
 23. A method according to claim 22, wherein, the ratio of 1,4-dioxane to THF is about 1:4.
 24. A method according to claim 21, wherein the base comprises potassium t-butoxide.
 25. A method according to claim 21, wherein the mole ratio of the base to methyl propionate is from about 3:1 to about 1:1.
 26. A method according to claim 21, wherein the nonpolar organic solvent used in Step (iii) comprises toluene.
 27. A method according to claim 21, wherein the nonpolar organic solvent in Step (v) comprises toluene.
 28. A method according to claim 21, wherein the nonpolar solvent in Step (iii) is the same nonpolar solvent used in Step (v).
 29. A method according to claim 21, wherein the yield of oltipraz is greater than 21% based on the amount of pyrazine-2-carboxylic acid methyl ester.
 30. A composition comprising oltipraz prepared by the process of claim
 21. 31. A method of making a purified oltipraz composition, comprising the steps of: (i) reacting a quantity of pyrazine-2-carboxylic acid methyl ester with methyl propionate to form methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, wherein the reaction is carried out in the presence of a base comprising potassium t-butoxide or sodium pentanoate, and a solvent comprising 1,4-dioxane in tetrahydrofuran (THF), wherein the ratio of 1,4-dioxane in the THF is at least about 1:5; (ii) adding at least one aqueous liquid to quench the reaction in Step (i); (iii) adding a nonpolar organic solvent and an ionic salt, thereby forming a composition comprising an aqueous component and an organic component, wherein the organic component comprises the nonpolar solvent and methyl 2-methyl-3-oxo-3-(pyrazin-2-yl) propanoate; (iv) separating the organic component from the aqueous component; (v) reacting the methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate ester in the organic component with P₂S₅ in the presence of a nonpolar organic solvent to form 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione (oltipraz); (vi) quenching the reaction with an aqueous composition to yield a composition comprising (i) an aqueous component and (ii) an organic component that comprises oltipraz; (vii) separating the organic component comprising oltipraz to yield an organic composition comprising oltipraz in the nonpolar organic solvent; (viii) reducing the amount of solvent in the organic composition; (ix) precipitating the oltipraz in the organic composition to yield a composition comprising oltipraz; (x) separating the precipitated oltipraz; and (xi) purifying the separated oltipraz precipitate.
 32. A method according to claim 31, wherein the ratio of 1,4-dioxane to THF in Step (i) is from about 1:5 to about 1:3.
 33. A method according to claim 31, wherein step (viii) comprises evaporating at least a portion of the solvent in the organic composition, step (ix) comprises adding to the organic composition a solvent that causes the oltipraz to precipitate, and step (x) comprises separating the oltipraz by filtering.
 34. A method according to claim 31, wherein the step of purifying comprises recrystallization, and excludes purification that comprises chromatographic separation.
 35. A method of making a composition comprising methyl 2-methyl-3-oxo-3-(pyrazin-2-yl), comprising the step of (i) reacting a quantity of pyrazine-2-carboxylic acid methyl ester with methyl propionate to form methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate, wherein the reaction is carried out in the presence of a base comprising potassium t-butoxide or sodium pentanoate, and a solvent comprising 1,4-dioxane in tetrahydrofuran (THF), wherein the ratio of 1,4-dioxane in the THF is at least about 1:5.
 36. A method according to claim 35, wherein the yield of methyl 2-methyl-3-oxo-3-(pyrazin-2-yl)propanoate is greater than 80% based on the amount of pyrazine-2-carboxylic acid methyl ester.
 37. A method according to claim 35, further comprising the step of adding at least one aqueous liquid to quench the reaction in Step (i), thereby forming a composition comprising an organic component and an aqueous component.
 38. A method according to claim 37, further comprising the step of separating the organic component from the aqueous component to yield a composition comprising the organic component.
 39. A composition made by the method of claim
 37. 40. A composition made by the method of claim
 38. 